Effect Of Qizhizhoufei Granules On Pulmonary Micro-angiogenesis Of Chromatic Obstructive Pulmonary Disease (COPD) Model Rats

Objective:To investigate prelim nary discussion about the treating effect of Qizhizhoufeigranules on COPD model rats and its function mechanism with respect to micro-angiogenesisby observing the effect of Qizhizhoufei granules on pathologic morphology of COPD modelrats’ pulmonary tissues,protein expression and transcriptional level of vascular endothelialgrowth factor (VEGF),and microvessel density (MVD).Methods:60Wistar rats of SPF level were randomly divided into six groups:blank group,COPD model group,positive control group and high,middle and low dose groups ofQizhizhoufei granules (ten in each group).The COPD rat model was built with compoundfactors:cigarette smoke plus intratracheal instillation of lipopolysaccharide (LPS).From15thday of the experiment,the blank group and COPD model group began accepting the perfusionwith normal saline,positive control group with roflumilast solution,and high,middle and lowdose groups of Qizhizhoufei granules with suspensions of different concentrations.Intervention treatment was administered for4weeks in all.Animals were put to death on the43rdday and samples were collected.HE staining,MOSSON staining,toluidine blue stainingand transmission electron microscope (TEM) were used to observe the pathomorphism aswell as change in ultra microstructure.The VEGF expression and MVD of rats’ pulmonarytissue was detected with immunohistochemical SABC,VEGF transcription level withRT-PCR,and SP-A content expression with immunofluorescent staining.Results:1. Effect on pathomorphology of pulmonary tissue: model group adopting HE stainingrevealed highly expanded pulmonary alveoli in some areas of the pulmonary tissue,narrowedand even ruptured alveolar interval,forced close of capillaries on alveolar wall,symptoms ofsevere emphysema, spasm and constriction of the bronchioles,and narrowed lumen, thickenedand sclerous wall of interstitial small vessels,and focal invasion of large number oflymphocytes and macrophagocytes on the bronchiole wall and around and between bloodvessels.As for positive control group,the invasion of inflammatory cells was relieved to somedegree,but not significantly.In the group administered high dose of Qizhizhoufei granules,theinvasion of inflammatory cells was greatly stopped and thickening and sclerosis of the small vascular wall was observed.The treatment also improved the spasm of bronchioles.Massonstaining showed the model group,when compared with blank group,to have markedlyincreased deposition of collagenous fiber in between pulmonary alveoli,bronchia,bronchioles,and the surrounding areas.In accordance with the results of TEM,within thecytoplasm,there were swollen mitochondria,broken or disappeared mitochondrial cristae,greatly reduced lamellar bodies with local focal dissolution,and significantly decreased villiat the edge of plasmalemma.In comparison with model group,the high dose group ofQizhizhoufei granules displayed relatively complete.Type II alveolar epithelial cells,significant mitochondrial swelling,reduced vacuolation in lamellar bodies,and greatlyincreased number of micro-villi.2. MVD:the model group had markedly increased MVD when compared with the blankgroup,and the difference was of statiscal significance (P<0.01).In comparison with modelgroup,the positive control group had a relatively decreased MVD and the difference was ofstiastical significance (P<0.05),and the high dose group of Qizhizhoufei granules hadsignificantly reduced MVD and the difference was of statiscal significance (P<0.05).3. VEGF:after the immumohistochemical staining,compared with blank group,the modelgroup revealed noticeably enhanced VEGF expression and the significance was of statiscalsignificance (P<0.01).The positive control group’s VEGF expression was between that ofmodel group and group treated with traditional Chinese medicine,but the difference withmodel group was of no statiscal significance (P>0.05).The high dose group of Qizhizhoufeigranules,compared with model group,showed obviously reduced VEGF level and thedifference was of statiscal significance (P<0.01).The VEGF mRNA expression of rats’ pulmonary tissue in model group wasstrengthened when compared with that in blank group,and the difference was of statisticalsignificance (P<0.05).In comparison with the model group,the high and middle dose groupsof Qizhizhoufei granules had reduced VEGF mRNA expression and the difference was alsostatistically significant (P<0.05).4. SP-A:in the model group,the SP-A fluorescent expression of rat’s pulmonary tissuewas significantly reduced while the SP-A mean alveolar positive cell number wassignificantly different from that in blank group (P<0.01).Compared with the model group,the positive control group displayed increased mean alveolar positive cell number,and thedifference was statistically significant (P<0.05);while the high and middle dose groups ofQizhizhoufei granules revealed strengthened SP-A fluorescent expression,and markedlyincreased SP-A mean alveolar positive cell number,and the difference was of statisticalsignificance (P<0.01).Conclusion:1. The experiment succeeded in copying COPD rat model through cigarette smoke incombination with intratracheal instillation of LPS.2. MVD and VEGF were proved to be critical for the micro-angiogenesis in pulmonarytissue of COPD rats.The study about them is helpful for the further exploration intopathogenesis of COPD and supply of academic basis for clinical treatment.3. The possible mechanism to account for the effective treatment of COPD withQizhizhoufei granules is the inhibition of VEGF expression and MVD so as to restrict thepathological micro-angiogenesis and cure COPD.